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Substantial economic losses in animal agriculture result from animals experiencing heat stress (HS). Pigs are especially susceptible to HS, resulting in reductions in growth, altered body composition, and compromised substrate metabolism. In this study, an artificial high-intensity sweetener and capsaicin (CAPS-SUC; Pancosma, Switzerland) were supplemented in combination to mitigate the adverse effects of HS on pig performance. Forty cross-bred barrows (16.2 ± 6 kg) were assigned to one of five treatments: thermal neutral controls (TN) (22 ± 1.2 °C; 38%-73% relative humidity) with ad libitum feed, HS conditions with ad libitum feed with (HS+) or without (HS-) supplementation, and pair-fed to HS with (PF+) or without supplementation (PF-). Pigs in heat-stressed treatments were exposed to a cyclical environmental temperature of 12 h at 35 ± 1.2 °C with 27%-45% relative humidity and 12 h at 30 ± 1.1 °C with 24%-35% relative humidity for 21 d. Supplementation (0.1 g/kg feed) began 7 d before and persisted through the duration of environmental or dietary treatments (HS/PF), which lasted for 21 d. Rectal temperatures and respiration rates (RR; breaths/minute) were recorded thrice daily, and feed intake (FI) was recorded daily. Before the start and at the termination of environmental treatments (HS/PF), a muscle biopsy of the longissimus dorsi was taken for metabolic analyses. Blood samples were collected weekly, and animals were weighed every 3 d during treatment. Core temperature (TN 39.2 ± 0.02 °C, HS- 39.6 ± 0.02 °C, and HS+ 39.6 ± 0.02 °C, P0.001) and RR (P0.001) were increased in both HS- and HS+ groups, but no difference was detected between HS- and HS+. PF- pigs exhibited reduced core temperature (39.1 ± 0.02 °C, P0.001), which was restored in PF+ pigs (39.3 ± 0.02 °C) to match TN. Weight gain and feed efficiency were reduced in PF- pigs (P0.05) but not in the PF+ or the HS- or HS+ groups. Metabolic flexibility was decreased in the HS- group (-48.4%, P0.05) but maintained in the HS+ group. CAPS-SUC did not influence core temperature or weight gain in HS pigs but did restore core temperature, weight gain, and feed efficiency in supplemented PF pigs. In addition, supplementation restored metabolic flexibility during HS and improved weight gain and feed efficiency during PF, highlighting CAPS-SUC's therapeutic metabolic effects.Heat stress reduces pig performance due to metabolic responses to heat. During heat stress, pigs lose the ability to metabolize fatty acids for energy and rely on carbohydrates to fuel growth. Evidence has shown that capsaicin, the active ingredient in chili peppers, interacts with heat-sensing receptors to protect against heat stress by preventing changes to metabolism. Artificial sweeteners can also preserve fat metabolism by inducing the secretion of metabolic regulatory hormones from the gut. This study examined a combination of capsaicin and artificial sweetener to restore growth and maintain metabolism during 3 wk of heat stress. As pigs often reduce their feed intake during heat stress, a group of pigs was feed restricted to match the reduced feeding observed in the heat-stressed pigs. Pigs given the feed supplement during heat stress maintained their metabolic flexibility, a measure of metabolic health. In agreement with previous short-term studies, the capsaicin and artificial sweetener supplement improved feed efficiency and weight gain in feed-restricted pigs. This study demonstrated that supplementation with capsaicin and artificial sweetener may prevent metabolic dysfunction during heat stress. This study also confirmed that supplementation with capsaicin and artificial sweetener does improve feed-restricted pigs’ growth and feed efficiency.

BACKGROUND Dietary calcium and phosphorus are required for bone and muscle development. Deficiencies of these macrominerals reduce bone mineral and muscle accretion potentially via alterations of mesenchymal stem cell (MSC) and satellite cell (SC) activities. OBJECTIVES With increasing interest in the role of early-life events on lifetime health outcomes, we aimed to elucidate the impact of dietary calcium and phosphorus, from deficiency through excess, on MSC and SC characteristics during neonatal development. METHODS Neonatal pigs [30 females, 1-d-old, 1.46 ± 0.04 kg body weight (BW)] were fed milk replacers for 16 d that were isonitrogenous and isocaloric with a consistent ratio of calcium to phosphorus, but either 25% deficient (calcium: 0.78%; phosphorus: 0.60%; CaPD), adequate (calcium: 1.08%; phosphorus: 0.84%; CaPA), or 25% in excess (calcium: 1.38%; phosphorus: 1.08%; CaPE) of calcium and phosphorus requirements based on sow-milk composition and extrapolation from NRC requirements for older pigs. BW and feed intake were recorded daily. Blood was collected for serum phosphorus, parathyroid hormone (PTH), and fibroblast growth factor 23 (FGF23) determination. Humeri were collected for MSC isolation and radii/ulnae bone were collected for analysis. Longissimus dorsi muscle was collected for SC isolation and analysis. RESULTS There was 4.6% increase in bone ash percentage in CaPE- versus CaPD-fed pigs (P

BACKGROUND Low-birth-weight (LBWT) neonates grow at a slower rate than their normal-birth-weight (NBWT) counterparts and may develop hypoglycemia postnatally. OBJECTIVE We investigated whether dietary lipid supplementation would enhance growth and improve glucose production in LBWT neonatal pigs. METHODS Twelve 3-d-old NBWT (1.606 kg) crossbred pigs were matched to 12 LBWT (1.260 kg) same-sex littermates. At 6 d of age, 6 pigs in each group were fed a low-energy (LE) or a high-energy (HE) isonitrogenous formula containing 5.2% and 7.3% fat, respectively. Body composition was assessed using dual-energy X-ray absorptiometry; plasma glucose and glycerol kinetics were assessed using stable isotope tracers. After killing, weights of skeletal muscles and visceral organs were measured. Data were analyzed by ANOVA for a 2 × 2 factorial design; temporal effects were investigated using repeated-measures analysis. RESULTS Lipid supplementation did not affect body weight of LBWT or NBWT pigs. However, liver and longissimus dorsi weights as a percentage of body weight were greater for pigs fed an HE diet than for those fed an LE diet (4.3% compared with 3.4% and 1.5% compared with 1.2%, respectively) but remained less for LBWT than for NBWT pigs (3.8% compared with 3.9% and 1.3% compared with 1.5%, respectively) (P

Study objectives were to determine the effects of rapamycin (Rapa) on biomarkers of metabolism and inflammation during acute heat stress (HS) in growing pigs. Crossbred barrows (n = 32; 63.5 ± 7.2 kg body weight [BW]) were blocked by initial BW and randomly assigned to 1 of 4 environmental-therapeutic treatments: 1) thermoneutral (TN) control (n = 8; TNCon), 2) TN and Rapa (n = 8; TNRapa), 3) HS control (n = 8; HSCon), or 4) HS and Rapa (n = 8; HSRapa). Following 6 d of acclimation to individual pens, pigs were enrolled in two experimental periods (P). During P1 (10 d), pigs were fed ad libitum and housed in TN conditions (21.3 ± 0.2°C). During P2 (24 h), HSCon and HSRapa pigs were exposed to constant HS (35.5 ± 0.4°C), while TNCon and TNRapa pigs remained in TN conditions. Rapamycin (0.15 mg/kg BW) was orally administered twice daily (0700 and 1800 hours) during both P1 and P2. HS increased rectal temperature and respiration rate compared to TN treatments (1.3°C and 87 breaths/min, respectively; P < 0.01). Feed intake (FI) markedly decreased in HS relative to TN treatments (64%; P < 0.01). Additionally, pigs exposed to HS lost BW (4 kg; P < 0.01), while TN pigs gained BW (0.7 kg; P < 0.01). Despite marked changes in phenotypic parameters caused by HS, circulating glucose and blood urea nitrogen did not differ among treatments (P > 0.10). However, the insulin:FI increased in HS relative to TN treatments (P = 0.04). Plasma nonesterified fatty acids (NEFA) increased in HS relative to TN treatments; although this difference was driven by increased NEFA in HSCon compared to TN and HSRapa pigs (P < 0.01). Overall, circulating white blood cells, lymphocytes, and monocytes decreased in HS compared to TN pigs (19%, 23%, and 33%, respectively; P ≤ 0.05). However, circulating neutrophils were similar across treatments (P > 0.31). The neutrophil-to-lymphocyte ratio (NLR) was increased in HS relative to TN pigs (P = 0.02); however, a tendency for reduced NLR was observed in HSRapa compared to HSCon pigs (21%; P = 0.06). Plasma C-reactive protein tended to differ across treatments (P = 0.06) and was increased in HSRapa relative to HSCon pigs (46%; P = 0.03). Circulating haptoglobin was similar between groups. In summary, pigs exposed to HS had altered phenotypic, metabolic, and leukocyte responses; however, Rapa administration had limited impact on outcomes measured herein.

Heat stress can negatively impact pig health and performance but the effects of heat stress on skeletal muscle mitochondrial function are largely unknown. We hypothesized that mitochondrial function and capacity would be impaired in heat stressed (HS) compared to thermoneutral (TN) pigs but mitochondrially-targeted coenzyme Q (MitoQ) supplementation would rescue the impairment. Oxidative portions of the semitendinosus muscle were evaluated from TN and HS gilts receiving no supplementation (CON) or MitoQ for 2 d prior to and during the 24h environmental heat treatment (n = 8 per group). Mitochondrial oxidative phosphorylation (P) and electron transfer (E) capacities were determined via high resolution respirometry and mitochondrial volume density and function were quantified by citrate synthase (CS) and cytochrome c oxidase activities, respectively. Data were analyzed using linear models in SAS v9.4 with fixed effects of heat, MitoQ treatment (trt), and heat×trt interaction. There were trends for the interaction of trt and heat (P≤0.1) on integrative (per mg tissue) and intrinsic (relative to CS) P with complexes I and II (PCI+II), maximum noncoupled E (ECI+II), and E with complex II only (ECII), in which all measures were greater in HS-MitoQ than TN-MitoQ (P≤0.03), but measures did not differ due to HS in CON pigs. The contribution of leak to total E (flux control ratio, FCRLeak) was lesser in HS-MitoQ than HS-CON, TN-CON, and TN-MitoQ (P≤0.02). The FCRPCI was greater (P≤0.05) while the FCRPCI+II was lesser (P=0.01) in TN compared to HS pigs. Finally, the FCRPCI+II was greater (P=0.02) while the FCRECII tended to be lesser (P=0.09) for CON than MitoQ pigs. Neither mitochondrial volume density nor function were affected by HS or MitoQ supplementation. In total, these data indicate improved mitochondrial capacities following heat stress in pigs receiving MitoQ but no difference in mitochondrial capacities in unsupplemented, HS pigs.

Adverse weather conditions are a large constraint to maximizing farm animal productivity. Heat stress, in particular, compromises almost every metric of animal agriculture profitability. Suboptimal production during HS was traditionally thought to result from hypophagia. However, independent of inadequate nutrient consumption, HS affects a plethora of endocrine, physiological, metabolic, circulatory, and immunological variables. Mounting evidence suggest that direct effects of HS originating at the gastrointestinal tract precede the observed effects on the aforementioned systems. Heat stress compromises intestinal barrier integrity causing the appearance of luminal contents, e.g. endotoxin, in circulation. Endotoxin stimulates both a classic immune response with local and systemic inflammatory reactions as well as directly acting on numerous organs and tissues. Once activated, leukocytes switch from oxidative phosphorylation to aerobic glycolysis where the glucose requirement of an intensely triggered immune system can exceed 2 kg/d in a lactating dairy cow. Whole body metabolic adjustments are primarily characterized by increased basal and stimulated circulating insulin, increased hepatic glucose output, decreased adipose tissue mobilization, and decreased skeletal muscle flexibility characterized by a reliance on glucose as a fuel substrate rather than lipid. Ultimately, the metabolic and physiological consequences of heat stress share a similar phenotype with immune challenges. Describing the physiology and mechanisms that underpin how HS jeopardizes animal performance is critical for developing approaches to ameliorate current production issues and requisite for generating future strategies (genetic, managerial, nutritional, and pharmaceutical) aimed at optimizing animal well-being, and improving the sustainable production of high-quality protein for human consumption.

Heat stress (HS) diminishes animal production, reducing muscle growth and increasing adiposity, especially in swine. Excess heat creates a metabolic phenotype with limited lipid oxidation that relies on aerobic and anaerobic glycolysis as a predominant means of energy production, potentially reducing metabolic rate. To evaluate the effects of HS on substrate utilization and energy expenditure, crossbred barrows (15.2 &plusmn, 2.4 kg) were acclimatized for 5 days (22 &deg, C), then treated with 5 days of TN (thermal neutral, 22 &deg, C, n = 8) or HS (35 &deg, C, n = 8). Pigs were fed ad libitum and monitored for respiratory rate (RR) and rectal temperature. Daily energy expenditure (DEE) and respiratory exchange ratio (RER, CO2:O2) were evaluated fasted in an enclosed chamber through indirect calorimetry. Muscle biopsies were obtained from the longissimus dorsi pre/post. HS increased temperature (39.2 &plusmn, 0.1 vs. 39.6 &plusmn, 0.1 &deg, C, p &lt, 0.01) and RER (0.91 &plusmn, 0.02 vs. 1.02 &plusmn, 0.02 VCO2:VO2, p &lt, 0.01), but decreased DEE/BW (68.8 &plusmn, 1.7 vs. 49.7 &plusmn, 4.8 kcal/day/kg, p &lt, 0.01) relative to TN. Weight gain (p = 0.80) and feed intake (p = 0.84) did not differ between HS and TN groups. HS decreased muscle metabolic flexibility (~33%, p = 0.01), but increased leucine oxidation (~35%, p = 0.02) compared to baseline values. These data demonstrate that HS disrupts substrate regulation and energy expenditure in growing pigs.

Objectives were to determine the effects of mitoquinol (MitoQ) on performance, metabolism, and inflammation during acute heat stress (HS) in growing pigs. Crossbred barrows (n=32; 59±1 kg BW) where blocked by BW and randomly assigned to 1 of 4 therapeutic-environmental treatments: 1) thermoneutral (TN) control (n=8; TNCtl), 2) TN and MitoQ (n=8; TNMitoQ), 3) HS control (n=8; HSCtl), or 4) HS and MitoQ (n=8; HSMitoQ). The trial consisted of two experimental periods (P). During P1 (2d), pigs were fed ad libitum and housed in TN conditions (20.6±0.1°C). During P2 (24h), HSCtl and HSMitoQ pigs were exposed to continuous HS (35.2±0.03°C); while TNCtl and TNMitoQ remained in TN conditions. Mitoquinol was orally administered twice daily (0700 and 1800 h; 40 mg/d) during P1 and P2. Pigs exposed to HS had increased rectal temperature, skin temperature, and respiration rate (1.46°C, 6.79°C, and 101 bpm, respectively; P< 0.01) compared to their TN counterparts. Acute HS markedly decreased feed intake (67%; P< 0.01). Additionally, HS pigs lost BW compared to their TN counterparts (-4.7 vs. +1.6 kg, respectively; P< 0.01); however, the reduction in BW was less severe in HSMitoQ compared to HSCtl pigs (-3.85 vs. -5.50 kg, respectively; P< 0.01). Circulating glucose increased in HSMitoQ relative to HSCtl pigs (15%; P=0.04). Non-esterified fatty acid (NEFA) concentrations were increased in HS compared to TN pigs (P< 0.01), although this difference was influenced by increased NEFA in HSCtl relative to HSMitoQ pigs (251 vs. 142 μEq/L; P< 0.01). Insulin:feed intake tended to increase in HS relative to TN pigs (P=0.09). Overall, no differences in blood urea nitrogen or cell blood counts were observed across treatments (P >0.10). In conclusion, acute HS exposure negatively altered animal performance and metabolism; however, administering MitoQ appeared to ameliorate the HS response.

Objectives were to determine the effects of rapamycin on biomarkers of metabolism and inflammation during acute heat stress (HS) in growing pigs. Crossbred barrows (n=32; 63.5±0.8 BW) where blocked by BW and randomly assigned to 1 of 4 therapeutic-environmental treatments: 1) thermoneutral (TN) control (n=8; TNCtl), 2) TN and rapamycin (n=8; TNRapa), 3) HS control (n=8; HSCtl), or 4) HS and rapamycin (n=8; HSRapa). The trial consisted of 2 experimental periods (P). During P1 (10d), pigs were fed ad libitum and housed in TN conditions (21.3±0.01°C). During P2 (24h), HSCtl and HSRapa pigs were exposed to constant HS (35.5±0.1°C); while TNCtl and TNRapa remained in TN conditions. Rapamycin (0.15 mg/kg BW) was orally administered twice daily (0700 and 1800 h) during P1 and P2. HS increased rectal temperature, skin temperature, and respiration rate compared to TN counterparts (1.28°C, 8.68°C, and 87 bpm, respectively; P< 0.01). Feed intake (FI) markedly decreased in HS relative to TN treatments (64%; P< 0.01). Additionally, pigs exposed to HS lost BW (4 kg; P< 0.01), while TN pigs gained BW (0.7 kg; P< 0.01). Overall, circulating white blood cells decreased in HS compared to TN pigs (19%; P=0.01). Circulating neutrophils did not differ across treatments; however, lymphocytes, monocytes, and basophils decreased in HS relative to TN pigs (23, 33, and 38%, respectively; P≤0.07). Despite marked changes in phenotypic parameters, circulating glucose and blood urea nitrogen did not differ among treatments (P >0.10). However, insulin:FI increased in HS relative to TN treatments (P=0.04). Plasma non-esterified fatty acids (NEFA) increased in HS relative to TN treatments; although this difference was driven by increased NEFA in HSCtl compared to TN and HSRapa pigs (P< 0.01). In summary, pigs exposed to HS had altered phenotypic, metabolic, and leukocyte responses; however, rapamycin administration had little to no effect on any of the variables measured.

Inflammation appears to be a predisposing factor and key component of hepatic steatosis in a variety of species. Objectives were to evaluate effects of inflammation [induced via intravenous lipopolysaccharide (LPS) infusion] on metabolism and liver lipid content in experimentally induced hyperlipidemic lactating cows. Cows (765 ± 32 kg of body weight; 273 ± 35 d in milk) were enrolled in 2 experimental periods (P); during P1 (5 d), baseline data were obtained. At the start of P2 (2 d), cows were assigned to 1 of 2 treatments: (1) intralipid plus control (IL-CON; 3 mL of saline; n = 5) or (2) intralipid plus LPS (IL-LPS; 0.375 μg of LPS/kg of body weight; n = 5). Directly following intravenous bolus (saline or LPS) administration, intralipid (20% fat emulsion) was intravenously infused continuously (200 mL/h) for 16 h to induce hyperlipidemia during which feed was removed. Blood samples were collected at -0.5, 0, 4, 8, 12, 16, 24, and 48 h relative to bolus administration, and liver biopsies were obtained on d 1 of P1 and at 16 and 48 h after the bolus. By experimental design (feed was removed during the first 16 h of d 1), dry matter intake decreased in both treatments on d 1 of P2, but the magnitude of reduction was greater in LPS cows. Dry matter intake of IL-LPS remained decreased on d 2 of P2, whereas IL-CON cows returned to baseline. Milk yield decreased in both treatments during P2, but the extent and duration was longer in LPS-infused cows. Administering LPS increased circulating LPS-binding protein (2-fold) at 8 h after bolus, after which it markedly decreased (84%) below baseline for the remainder of P2. Serum amyloid A concentrations progressively increased throughout P2 in IL-LPS cows (3-fold, relative to controls). Lipid infusion gradually increased nonesterified fatty acids and triglycerides in both treatments relative to baseline (3- and 2.5-fold, respectively). Interestingly, LPS infusion blunted the peak in nonesterified fatty acids, such that concentrations peaked (43%) higher in IL-CON compared with IL-LPS cows and heightened the increase in serum triglycerides (1.5-fold greater relative to controls). Liver fat content remained similar in IL-LPS relative to P1 at 16 h; however, hyperlipidemia alone (IL-CON) increased liver fat (36% relative to P1). No treatment differences in liver fat were observed at 48 h. In IL-LPS cows, circulating insulin increased markedly at 4 h after bolus (2-fold relative to IL-CON), and then gradually decreased during the 16 h of lipid infusion. Inducing inflammation with simultaneous hyperlipidemia altered the characteristic patterns of insulin and LPS-binding protein but did not cause fatty liver.

Unfavorable weather conditions are one of the largest constraints to maximizing farm animal productivity. Heat stress (HS), in particular, compromises almost every metric of profitability and this is especially apparent in the grow-finish and reproductive aspects of the swine industry. Suboptimal production during HS was traditionally thought to result from hypophagia. However, independent of inadequate nutrient consumption, HS affects a plethora of endocrine, physiological, metabolic, circulatory, and immunological variables. Whether these changes are homeorhetic strategies to survive the heat load or are pathological remains unclear, nor is it understood if they temporally occur by coincidence or if they are chronologically causal. However, mounting evidence suggest that the origin of the aforementioned changes lie at the gastrointestinal tract. Heat stress compromises intestinal barrier integrity, and increased appearance of luminal contents in circulation causes local and systemic inflammatory responses. The resulting immune activation is seemingly the epicenter to many, if not most of the negative consequences HS has on reproduction, growth, and lactation. Interestingly, thermoregulatory and production responses to HS are only marginally related. In other words, increased body temperature indices poorly predict decreases in productivity. Further, HS induced malnutrition is also a surprisingly inaccurate predictor of productivity. Thus, selecting animals with a "heat tolerant" phenotype based solely or separately on thermoregulatory capacity or production may not ultimately increase resilience. Describing the physiology and mechanisms that underpin how HS jeopardizes animal performance is critical for developing approaches to ameliorate current production issues and requisite for generating future strategies (genetic, managerial, nutritional, and pharmaceutical) aimed at optimizing animal well-being, and improving the sustainable production of high-quality protein for human consumption.

Muscle hypertrophy is limited in low birth weight (LBWT) neonates, suggesting a reduction in protein synthesis and increased protein degradation. Sixteen pairs of one-day-old normal birth weight (NBWT) and LBWT littermates ( n = 16) were euthanized, and the longissimus dorsi (LD) was sampled for protein abundance and kinase phosphorylation profile measures. Eukaryotic initiation factor (eIF)4E and eIF4G abundance, and assembly of the active eIF4E-eIF4G complex, was less for LBWT than for NBWT pig muscles. Similarly, eIF3f abundance was reduced in the muscle of LBWT compared with NBWT pigs and was associated with diminished ribosomal protein S6 kinase 1 phosphorylation. This decrease was linked to a lower phosphorylation of programmed cell death protein 4 (PDCD4) in LBWT pig muscle. By contrast, PDCD4 abundance was greater in the muscle of the LBWT than NBWT group, suggesting lower release and availability of eIF4A from the PDCD4-eIF4A complex. Moreover, protein abundance of eIF4A was lower in LBWT muscle, which is expected to further impair the formation of eIF4F translation initiation complex. Microtubule-associated light chain 3 (LC3) II to total LC3 ratio was greater in LBWT LD lysates, yet P62 abundance was similar between the two groups, suggesting no difference in autophagy. Muscle atrophy F-box (atrogin-1) abundance was less in LBWT LD lysates, suggesting decreased degradation through the ubiquitin-proteasome system. In conclusion, limited eIF4F subunit abundance and downregulated translation initiation are plausible mechanisms for diminished muscle growth in LBWT compared with NBWT neonatal pigs.NEW & NOTEWORTHY We demonstrated that eukaryotic initiation factor (eIF)4E, eIF4G, and eIF4A abundance, and assembly of the active eIF4E-eIF4G complex, were reduced in low birth weight (LBWT) compared with normal birth weight pig muscle. In contrast, our data indicated that protein degradation signaling does not seem to affect protein turnover in LBWT pig muscle. Thus, downregulated translation initiation is likely the key contributor that predisposes LBWT neonatal pigs to slower postnatal muscle growth.

Study objectives were to determine the effects of zinc (Zn) amino acid complex (Availa Zn, Zinpro Corporation, Eden Prairie, MN) on metabolism, biomarkers of leaky gut, and inflammation during and following heat stress (HS) and nutrient restriction. Crossbred gilts (n = 50; 50 ± 2 kg BW) were blocked by initial BW and randomly assigned to one of five treatments: 1) thermoneutral (TN) and ad libitum fed a control diet (TNCtl), 2) TN and pair-fed a control diet (PFCtl), 3) TN and pair-fed a Zn-supplemented diet (PFZn), 4) HS and ad libitum fed a control diet (HSCtl), and 5) HS and ad libitum fed a Zn-supplemented diet (HSZn). The study consisted of 3 experimental periods (P): during P1 (7 d), all pigs were fed their respective diets ad libitum and housed in TN conditions (20.84 ± 0.03 °C, 47.11 ± 0.42% relative humidity). During P2 (7 d), HSCtl and HSZn pigs were exposed to progressive cyclical HS conditions (27 to 30 °C, 41.9 ± 0.5% relative humidity), while TNCtl, PFCtl, and PFZn pigs remained in TN conditions and were fed ad libitum or pair-fed to their respective HSCtl and HSZn counterparts. During P3 (5 d; "recovery phase"), all pigs were housed in TN conditions and fed ad libitum. Pigs exposed to HS had overall increased rectal temperature, skin temperature, and respiration rate (0.33 °C, 3.76 °C, and 27 bpm, respectively; P 0.10). In summary, Zn reduced TNFα (regardless of HS), and the stimulatory effect of HS on insulin secretion is amplified during HS recovery.

Prolonged heat stress represents a continuing threat to human health and agricultural production. Despite the broad, negative impact of prolonged hyperthermia little is known about underlying pathological mechanisms leading to negative health outcomes, which has limited the development of etiological interventions and left clinicians and producers with only cooling and rehydration strategies. The purpose of this investigation was to determine the extent to which prolonged environment-induced hyperthermia altered autophagy in oxidative skeletal muscle in a large animal model, serving the dual purpose of accurately modeling human physiology as well as agricultural production. We hypothesized that prolonged hyperthermia would induce autophagy in skeletal muscle, independent of the accompanying caloric restriction. To test this hypothesis pigs were treated as follows: thermoneutral (20 °C), heat stress (35 °C), or were held under thermoneutral conditions but pair-fed to the heat stress group for seven days. Upon euthanasia the red portion of the semitendinosus was collected. We found that prolonged hyperthermic exposure increased oxidative stress without a corresponding change in antioxidant enzyme activities. Hyperthermia prevented initiation of autophagy despite increased markers of nucleation, elongation and autophagosome formation. However, p62 relative protein abundance, which is inversely correlated with autophagic degradation, was strongly increased suggesting suppressed degradation of autophagosomes. Markers of mitophagy and mitochondrial abundance were largely similar between groups. These data indicate that faulty autophagy plays a key role in hyperthermic muscle dysfunction.

Heat stress (HS) causes morbidities and mortalities, in part by inducing organ-specific injury and dysfunction. Further, HS markedly reduces farm animal productivity, and this is especially true for lean tissue accretion. The purpose of this investigation was to determine the extent to which short-term HS caused muscle dysfunction in skeletal muscle. We have previously found increased free radical injury in skeletal muscle following 24 h of HS. Thus, we hypothesized that HS would lead to apoptosis, autophagy, and decreased mitochondrial content in skeletal muscle. To test this hypothesis, crossbred gilts were divided into 3 groups ( = 8/group): thermal neutral (TN: 21°C), HS (37°C), and pair-fed thermal neutral (PFTN: feed intake matched with heat-stressed animals). Following 12 h of treatment, animals were euthanized and red (STR) and white (STW) portions of the semitendinosus were recovered. Heat stress did not alter intracellular signaling in STW. In STR, the oxidative stress marker malondialdehyde protein and concentration were increased in HS ( = 0.007) compared to TN and PFTN, which was matched by an inadequate antioxidant response, including an increase in superoxide dismutase (SOD) I ( = 0.03) and II relative protein abundance ( = 0.008) and total SOD activity ( = 0.02) but a reduction ( = 0.006) in catalase activity in HS compared to TN. Further, B-cell lymphoma 2-associated X protein ( = 0.02) and apoptotic protease activating factor 1 ( = 0.01) proteins were increased by HS compared to TN and PFTN. However, caspase 3 activity was similar between groups, indicating a lack of apoptotic execution. Despite increased initiation, autophagy appeared to be inhibited by HS as the microtubule-associated protein A/B light chain 3 II/I ratio and mitofusin-2 proteins were decreased ( < 0.03) and sequestosome 1(p62) protein abundance was increased ( = 0.001) in HS compared to TN and PFTN. Markers of mitochondrial content cytochrome c, cytochrome c oxidase IV, voltage-dependent anion channel, pyruvate dehydrogenase, and prohibitins 1 were increased ( < 0.05) in HS compared to TN, whereas mitochondrial biogenesis and mitophagy markers were similar between groups. These data demonstrate that HS caused aberrant intracellular signaling, which may contribute to HS-mediated muscle dysfunction.

The objectives of this study were to elucidate the effects of a dirty environment and a challenge plus associated environmental contamination on pig growth performance, diet utilization efficiency, and gas emissions (CO, NH, CH, NO, and HS) from stored manure. Twenty-four weaned barrows, aged 31 d at initiation of the trial, were randomly allotted to 3 different treatments in a completely randomized design. Treatments were: pigs housed in cages with manure removed and cages washed daily (Clean); pigs housed in cages sprayed daily with manure slurry mixtures (Dirty); or pigs challenged with Typhimurium DT104 and housed in cages that were not washed, but manure was removed daily ( challenge). Rectal temperature, body weight, daily feed intake, manure output, manure composition, and gas emissions from stored manure were measured throughout the 24-d animal phase. The Dirty and challenge treatments were statistically compared to the Clean treatment to evaluate individual effects. Dirty housing tended to decrease ADG from d 1 to 24 ( = 0.06) but there were no other effects on pig performance compared with the Clean treatment. In contrast, a challenge was associated with a marked reduction in each of the measured indicators of pig performance. challenge increased the carbon to nitrogen ratio, ether extract, and lignin concentrations in excreted manure ( = 0.02, 0.01, 0.003, respectively), and increased manure and head space temperatures in manure tanks ( < 0.0001). Gas emissions from stored manure of pigs on the Dirty or treatments were increased for each of the measured gases as compared to the Clean treatment ( < 0.01) when expressed per unit of BW gain. When gas emissions from manure of pigs housed in the Dirty treatment were expressed per unit of manure volatile solids (VS), they were increased for NH, CH, and HS ( < 0.02). challenge was associated with increased emissions of CO, and NO and decreased emissions of HS per kilogram manure VS compared to the Clean treatment ( = 0.06, 0.03, 0.04, respectively). Collectively, these results indicated that a challenge and associated housing contamination caused depressed growth rate and increased manure gas emissions, while exposure to a Dirty environment slightly reduced growth performance and clearly increased manure gas emissions per unit of BW gain as compared to Clean control.

Pigs exposed to elevated ambient temperatures exhibit reduced daily gain, alterations in muscle and fat deposition, and decreased health. Negative aspects of gastrointestinal (GI) function, integrity, and permeability also occur. High-intensity sweeteners can ameliorate the negative effects of heat stress (HS) by increasing GI glucagon-like peptide-2 production while capsicum oleoresin has been shown to reduce inflammatory response. The effects of an artificial high-intensity sweetener and capsicum oleoresin (CAPS-SUC; TakTik X-Hit, Pancosma, Switzerland) on growth performance of pigs were examined. Forty-eight pigs (12 wk of age, 43.2 ± 4.3 kg) were assigned to six treatments: thermoneutral conditions (21 ± 1.1 °C; 40% to 70% relative humidity) fed ad libitum with (TN+) or without supplement (TN−), heat stress (35 ± 1 °C; 20% to 40% relative humidity) fed ad libitum with (HS+) or without supplement (HS−), and thermoneutral conditions pair-fed to HS intake with (PFTN+) or without supplement (PFTN−). Supplementation (0.1 g/kg feed) began 2 d prior to the 3-d environmental treatment period. Body weights (BWs) and blood samples were collected on days −1 and 3. Rectal temperature (RT) and respiration rate (RR) were measured thrice daily and the feed intake (FI) was recorded daily. Intestinal sections were collected for histology. Pigs in HS conditions exhibited increased RT (~1.2 °C) and RR (~2.7-fold) compared with TN and PFTN groups (P < 0.01). HS+ animals had increased RR when compared with HS− animals (P < 0.02). Heat stress decreased FI compared with TN. HS and PFTN decreased (P < 0.05) average daily gain compared with TN. Supplement did not alter the BW gain. HS and PFTN decreased (P < 0.05) Gain:Feed compared with TN during environmental treatment. Supplementation with CAPS–SUC increased Gain:Feed by 0.12 (P < 0.05). Circulating glucose concentrations tended to decrease in CAPS–SUC vs. non-supplemented HS and PFTN animals (P ≤ 0.1). Circulating insulin concentrations as well as monocyte count increased in HS compared with PFTN (P < 0.04) but did not differ from TN and likely linked to altered FI. CAPS–SUC increased basophil count (P < 0.02), irrespective of environment. Ileal villus height tended to decrease during HS and PFTN compared with TN (P < 0.08), indicating an effect of intake. Overall, CAPS–SUC supplementation increased pig feed efficiency and may improve immune response.

Heat stress continues to undermine efficient meat production and meat quality. It also jeopardizes human and animal health and wellbeing, regionalizes animal production, and threatens food security. Environmental models predict more frequent and severe heat waves, even in areas previously considered temperate indicating this problem will continue to have a progressively expanding, deleterious impact on agricultural productivity. Despite the broad, negative impact of heat stress little is known about underlying mechanisms leading to phenotypic outcomes. Because of its mass and energetic demands, skeletal muscle contributes greatly to regulation of systemic metabolism. We have discovered heat stress causes robust but transient oxidative stress and activation of apoptotic signaling in skeletal muscle in as little as two hours. Interestingly, these declined linearly through six hours concomitant with markers of increased autophagy and mitophagy, which would facilitate the removal of damaged mitochondria. Continued heating through 24 h causes a resumption of oxidative stress and autophagic dysfunction with an accumulation of autophagosomes and mitochondria. As mitochondrial injury and autophagic dysregulation appear to be key mediators of hyperthermic muscle dysfunction we propose a model that posits progressive mitochondrial injury leads to production of free radicals that overwhelms antioxidant systems and impairs autophagy facilitating accumulation of damaged, pro-oxidant mitochondria. Ultimately, these aforementioned changes may reduce efficient protein accretion. Our current work is focused on stimulating autophagy and protecting mitochondria during heat stress in an effort to maintain efficient muscle growth.

Heat stress (HS) poses a major threat to human health and agricultural production. Oxidative stress and mitochondrial dysfunction appear to play key roles in muscle injury caused by HS. We hypothesized that mitoquinol (MitoQ), would alleviate oxidative stress and cellular dysfunction in skeletal muscle during HS. To address this, crossbred barrows (male pigs) were treated with placebo or MitoQ (40 mg/d) and were then exposed to thermoneutral (TN; 20 °C) or HS (35 °C) conditions for 24 h. Pigs were euthanized following the environmental challenge and the red portion of the semitendinosus (STR) was collected for analysis. Unexpectedly, malondialdehyde concentration, an oxidative stress marker, was similar between environmental and supplement treatments. Heat stress decreased LC3A/B-I (p 0.05) and increased the ratio of LC3A/B-II/I (p 0.05), while p62 was similar among groups suggesting increased degradation of autophagosomes during HS. These outcomes were in disagreement with our previous results in muscle from gilts (female pigs). To probe the impact of biological sex on HS-mediated injury in skeletal muscle, we compared STR from these barrows to archived STR from gilts subjected to a similar environmental intervention. We confirmed our previous findings of HS-mediated dysfunction in muscle from gilts but not barrows. These data also raise the possibility that muscle from gilts is more susceptible to environment-induced hyperthermia than muscle from barrows.

At the onset of heat stress, cattle initiate a series of whole body adaptations in an effort to cope with and dissipate additional heat load. These include well-known physiological changes such as increased respiration rate and sweating rate and decreased feed intake. Environmentally induced hyperthermia in ruminants depresses production as a consequence of reduced feed intake but it is unclear how shifts in metabolism may further affect production performance and physiological acclimation. Our evidence indicates that cattle experiencing heat stress do not appear to engage metabolic and glucose-sparing adaptations consistent with their plane of nutrition. In this context, the liver is uniquely positioned to direct exogenously and endogenously derived nutrients for use by other metabolically active tissues such as the mammary gland and skeletal muscle. Despite the prominent role of the liver in whole-body metabolism, alterations in the molecular mechanisms leading to hepatic adaptation during heat challenge are unclear. We are using management tools and metabolic modifiers, such as bovine somatotropin, in an attempt to better understand and improve hepatic function during heat stress. Because a large proportion of an animal’s mass is comprised of skeletal muscle, alterations in skeletal muscle metabolism and function can have a profound impact on whole-animal energy metabolism and nutrient homeostasis especially during periods of stress. We have initiated studies to understand how hyperthermia influences the set points of several metabolic pathways within skeletal muscle. It appears that during heat stress bovine skeletal muscle experiences mitochondrial dysfunction leading to impaired cellular energy status. Finally, investigations into adipose tissue metabolism demonstrate impaired lipolytic functions likely due to a refractory nature to adrenergic stimuli. Taken together, this may have broad implications for the reduced production and heat intolerance seen during heat stress especially if tissue(s) are not able to make necessary contributions to whole-body energy homeostasis. Accurately understanding the biological mechanism(s) by which thermal stress reduces animal performance is critical for developing novel approaches (i.e. genetic, managerial and nutritional) to preserve growth and lactation especially given the critical importance of nutrients, such as glucose, to animal production and well being in these situations.

Basic principles governing skeletal muscle growth and development, from a cellular point of view, have been realized for several decades. Skeletal muscle is marked by the capacity for rapid hypertrophy and increases in protein content. Ultimately, skeletal muscle growth is controlled by 2 basic means: 1) myonuclear accumulation stemming from satellite cell (myoblast) proliferation and 2) the balance of protein synthesis and degradation. Each process underlies the rapid changes in lean tissue accretion evident during fetal and neonatal growth and is particularly sensitive to nutritional manipulation. Although multiple signals converge to alter skeletal muscle mass, postprandial changes in the anabolic hormone insulin link feed intake with enhanced rates of protein synthesis in the neonate. Indeed, a consequence of insulin-deficient states such as malnutrition is reduced myoblast activity and a net loss of body protein. A well-characterized mechanism mediating the anabolic effect of insulin involves the phosphatidylinositol 3-kinase (PI3K)-mammalian target of rapamycin (mTOR) signaling pathway. Activation of mTOR leads to translation initiation control via the phosphorylation of downstream targets. Modulation of this pathway by insulin, as well as by other hormones and nutrients, accounts for enhanced protein synthesis leading to efficient lean tissue accretion and rapid skeletal muscle gain in the growing animal. Dysfunctional insulin activity during fetal and neonatal stages likely alters growth through cellular and protein synthetic capacities.

Dietary protein intake during the neonatal period is critical for lifetime muscle development. Appropriate nutrition during the neonatal period may have lifelong impacts on animal growth and muscle development. We hypothesized that minor changes (25% ± adequacy) in dietary protein during the neonatal phase of life will impact future lean growth potential. The goal of this project was to determine the effects of a diet with a 25% deficiency (D), adequacy (A), and 25% excess in protein (E) on neonatal muscle hypertrophy. Dietary crude protein level in the adequate group (27%) was based upon sow’s milk composition and an extrapolation of NRC recommendations. Thirty, individually housed, 1-day-old piglets were divided into 3 groups and fed milk replacers with a 25% deficiency (D), adequacy (A), or 25% excess in protein (E) over a 17-day period. Body weight and feed intake were recorded daily, and a section of the Longissimus dorsi were collected for molecular and histological evaluation at the end of the experiment. Piglets fed E had significantly greater final body weight, (P

Heat-stressed pigs experience metabolic alterations, including altered insulin profiles, reduced lipid mobilization, and compromised intestinal integrity. This is bioenergetically distinct from thermal neutral pigs on a similar nutritional plane. To delineate differences in substrate preferences between direct and indirect (via reduced feed intake) heat stress effects, skeletal muscle fuel metabolism was assessed. Pigs (35.3 ± 0.8 kg) were randomly assigned to three treatments: thermal neutral fed ad libitum (TN; 21°C, n = 8), heat stress fed ad libitum (HS; 35°C, n = 8), and TN, pair-fed/HS intake (PF; n = 8) for 7 days. Body temperature (TB) and feed intake (FI) were recorded daily. Longissimus dorsi muscle was biopsied for metabolic assays on days −2, 3, and 7 relative to initiation of environmental treatments. Heat stress increased TBand decreased FI ( P < 0.05). Heat stress inhibited incomplete fatty acid oxidation and glucose oxidation ( P < 0.05). Metabolic flexibility decreased in HS pigs compared with TN and PF controls ( P < 0.05). Both phosphofructokinase and pyruvate dehydrogenase (PDH) activities increased in PF ( P < 0.05); however, TN and HS did not differ. Heat stress inhibited citrate synthase and β-hydroxyacyl-CoA dehydrogenase (β-HAD) activities ( P < 0.05). Heat stress did not alter PDH phosphorylation or carnitine palmitoyltransferase 1 abundance but reduced acetyl-CoA carboxylase 1 (ACC1) protein abundance ( P < 0.05). In conclusion, HS decreased skeletal muscle fatty acid oxidation and metabolic flexibility, likely involving β-HAD and ACC regulation.

Study objectives were to determine the effects of chromium (Cr) propionate (Cr propionate 0.04%; 0.5 g/kg of feed to deliver 200 parts per billion Cr/d; KemTRACE Cr, Kemin Industries, Inc., Des Moines, IA) on growth performance, metabolism, and health biomarkers in heat-stressed and nutrient-restricted pigs. Crossbred barrows (n = 96; 105 ± 1 kg BW) were enlisted in an experiment conducted in two replicates, blocked by initial BW, and randomly assigned to one of six dietary-environmental treatments: (i) thermoneutral (TN) and fed ad libitum a control diet (TNCtl), (ii) TN and fed ad libitum a Cr supplemented diet (TNCr), (iii) TN and pair-fed a control diet (PFCtl), (iv) TN and pair-fed a Cr supplemented diet (PFCr), (v) heat stress (HS) and ad libitum fed a control diet (HSCtl), or (vi) HS and ad libitum fed a Cr supplemented diet (HSCr). The study consisted of three experimental periods (P). During P0 (5 d), all pigs were housed in TN conditions (21.3 ± 0.1 °C, 56.8 ± 0.3% relative humidity [RH]) and fed the control diet ad libitum. During P1 (5 d), pigs were fed their respective dietary treatments ad libitum and kept in TN conditions. During P2 (35 d), HSCtl and HSCr-treated pigs were fed ad libitum and exposed to progressive cyclical HS conditions (27 to 31 °C, 50 ± 0.3% RH), while TNCtl, TNCr, PFCtl, and PFCr pigs remained in TN conditions and were fed ad libitum or pair-fed to their respective HSCtl and HSCr counterparts to eliminate the confounding effects of dissimilar feed intake. Overall, HS pigs had increased (P < 0.01) rectal temperature, skin temperature, and respiration rate (0.3 °C, 3.8 °C, and 32 breaths per minute, respectively) relative to TN pigs. Overall, HS decreased ADFI and ADG (20 and 21%, respectively; P < 0.01) compared with TN controls. Final BW tended to be increased in HSCr (2.7 kg, P = 0.06) compared with HSCtl pigs. Similarly, ADG tended to be increased during P2 in HSCr relative to HSCtl-treatment (0.77 vs. 0.72 kg/d; P = 0.10). There were no effects of Cr on most production parameters, but ADFI tended to be increased in Cr relative to Ctl-fed pigs (3.19 vs. 3.09 kg/d; P = 0.08). No effects of Cr supplementation were detected on circulating glucose, insulin, NEFA, cholesterol, triglycerides, or lipopolysaccharide binding protein. However, blood neutrophils were increased in HSCr (37%; P < 0.01) relative to HSCtl pigs. In summary, these results suggest Cr supplementation may benefit growth performance during HS.

Heat stress (HS) jeopardizes animal productivity and health. The intestinal barrier is sensitive to HS and heat-induced hyperpermeability plays a key role in its pathophysiology. However, the biology of recovery following HS is less understood. Thus, study objectives were to determine the temporal pattern of metabolic, inflammatory, and intestinal histological parameters during HS recovery. Female pigs (n = 32; 19.5 ± 0.5 kg BW) were sacrificed following exposure to 1 of 4 environmental treatments: 1) constant thermoneutral (TN) conditions (TNC; 24.2 ± 0.5°C), 2) no TN recovery post HS (0D), 3) 3 d of TN recovery post HS (3D), and 4) 7 d of TN recovery post HS (7D). The HS protocol was cyclical (33.6 ± 1.8 to 37.4 ± 2.1°C) and lasted for 3 d for all HS treatments. During the 3 d of HS, rectal temperature, skin temperature, and respiration rates were increased (1.3°C, 4.8°C, and 77 breaths/min, respectively; P < 0.01) and ADFI was decreased (27%; P < 0.01) compared to TNC pigs. Skin temperature tended to be decreased 0.6°C in 3D pigs during days 1-3 of recovery (P = 0.06) and was decreased 1.6 and 0.7°C during days 1-3 and 4-7 of recovery, respectively, in 7D pigs (P ≤ 0.03) compared to TNC. Relative to TNC pigs, ADFI remained 14% decreased during days 1-3 of recovery in both 3D and 7D pigs, and 17% decreased during days 4-7 in 7D pigs (P ≤ 0.01). Plasma glucose was decreased (10%; P = 0.03) for 0D and 3D relative to TNC pigs. Circulating lipopolysaccharide-binding protein was increased in 3D and 7D vs. TNC pigs (110 and 147%, respectively; P = 0.01) and tended to increase linearly with increasing recovery time (P = 0.08). Circulating tumor necrosis factor alpha was decreased (15%) in 0D pigs and increased linearly with advancing recovery time (P < 0.01). Jejunum and ileum villus height were reduced 17 and 11% in 0D vs. TNC pigs and increased linearly with progressive recovery time (P < 0.01). Jejunum and ileum mucosal surface areas were reduced 17 and 9% in 0D pigs and remained decreased in the jejunum while the ileum recovered to TNC levels by day 3 of recovery. Relative to TNC pigs, goblet cell area was similar in jejunum and colon of 0D pigs but was reduced in the ileum of 0D pigs and in jejunum, ileum, and colon of 3D and 7D relative to TNC pigs (P < 0.01). In summary, HS has deleterious effects on intestinal morphology that seem to improve with recovery time. In contrast, feed consumption remained suppressed and inflammatory biomarkers indicative of leaky gut increased following the heat load.

Study objectives were to determine the effects of zinc (Zn) amino acid complex (Availa Zn, Zinpro Corporation, Eden Prairie, MN) on metabolism, biomarkers of leaky gut, and inflammation during and following heat stress (HS) and nutrient restriction. Crossbred gilts (n = 50; 50 ± 2 kg BW) were blocked by initial BW and randomly assigned to one of five treatments: 1) thermoneutral (TN) and ad libitum fed a control diet (TNCtl), 2) TN and pair-fed a control diet (PFCtl), 3) TN and pair-fed a Zn-supplemented diet (PFZn), 4) HS and ad libitum fed a control diet (HSCtl), and 5) HS and ad libitum fed a Zn-supplemented diet (HSZn). The study consisted of 3 experimental periods (P): during P1 (7 d), all pigs were fed their respective diets ad libitum and housed in TN conditions (20.84 ± 0.03 °C, 47.11 ± 0.42% relative humidity). During P2 (7 d), HSCtl and HSZn pigs were exposed to progressive cyclical HS conditions (27 to 30 °C, 41.9 ± 0.5% relative humidity), while TNCtl, PFCtl, and PFZn pigs remained in TN conditions and were fed ad libitum or pair-fed to their respective HSCtl and HSZn counterparts. During P3 (5 d; “recovery phase”), all pigs were housed in TN conditions and fed ad libitum. Pigs exposed to HS had overall increased rectal temperature, skin temperature, and respiration rate (0.33 °C, 3.76 °C, and 27 bpm, respectively; P < 0.01). Relative to TN controls, HS decreased ADFI and ADG (28 and 35%, respectively; P < 0.05), but these variables were unaffected by dietary treatment. Additionally, circulating insulin did not differ between HS and TN pigs (P = 0.41), but was decreased in PF relative to TN pigs (P < 0.01). During recovery, no differences were observed in rectal temperature or respiration rate across treatments, but HSZn pigs had decreased skin temperature relative to TN, PF, and HSCtl pigs (P < 0.01). During P3, no Zn effects were observed in production parameters; however, PF pigs had increased ADFI and ADG relative to TN and HS treatments (P < 0.01). During P3, circulating insulin was increased in pigs that were HS relative to TN and PF pigs (75%, P < 0.05). Interestingly, tumor necrosis factor alpha (TNFα) levels were decreased during P3 (P = 0.04) in Zn relative to Ctl-fed pigs. Circulating lipopolysaccharide-binding protein was not different among periods (P > 0.10). In summary, Zn reduced TNFα (regardless of HS), and the stimulatory effect of HS on insulin secretion is amplified during HS recovery.

Heat stress (HS) affects numerous physiological processes including nutrient partitioning and lipid metabolism. Objectives of this study were to evaluate how acute HS affects lipid metabolism in subcutaneous adipose tissue of dairy cattle. Adipose tissue biopsies were performed on Holstein cows for bovine primary adipocyte isolation and cultured at either 42°C (HS) or 37°C (thermal neutral, TN). Adipocytes were incubated with increasing isoproterenol (ISO), and with increasing concentrations of insulin in the presence of ISO to evaluate changes in lipolysis. Incorporation of radioactive acetate into lipids was measured as an indicator of lipogenesis. Abundance and phosphorylation of several lipolytic and lipogenic proteins were also measured. Adipocytes exposed to HS had an elevated maximal response to ISO and were more sensitive to lipolytic stimulation by ISO compared with cells cultured at TN. Thermal treatment did not affect the antilipolytic effects of insulin in the presence of ISO. Lipogenesis measured as acetate incorporation was not altered by HS, but a temperature by insulin interaction was observed for the regulation of acetyl CoA carboxylase, such that the presence of insulin resulted in a reduction in phosphorylation of acetyl CoA carboxylase in adipocytes cultured at TN but not HS conditions. Results of this study demonstrate that acute HS has a direct effect on the regulation of lipolysis and the rate-limiting enzyme of lipogenesis in isolated bovine adipocytes.

We recently demonstrated that in utero heat stress (IUHS) alters future tissue accretion in pigs, but whether this is a conserved response among species, is due to the direct effects of heat stress (HS) or mediated by reduced maternal feed intake (FI) is not clear. Study objectives were to compare the quantity and rate of tissue accretion in rats exposed to differing in utero thermal environments while eliminating the confounding effect of dissimilar maternal FI. On d3 of gestation, pregnant Sprague-Dawley rats (189.0±5.9g BW) were exposed to thermoneutral (TN; 22.2±0.1°C; n=8), or HS conditions (cyclical 30 to 34°C; n=8) until d18 of gestation. A third group was pair-fed to HS dams in TN conditions (PFTN; 22.2±0.1°C; n=8) from d4 to d19 of gestation. HS increased dam rectal temperature (p=0.01; 1.3°C) compared to TN and PFTN mothers, and reduced FI (p=0.01; 33%) compared to TN ad libitum fed controls. Although litter size was similar (p=0.97; 10.9 pups/litter), pup birth weight was reduced (p=0.03; 15.4%) in HS compared to PFTN and TN dams. Two male pups per dam [n=8 in utero TN (IUTN); n=8 IUHS; n=8 in utero PFTN (IUPFTN)] were selected from four dams per treatment based on similar gestation length, and body composition was determined using dual-energy x-ray absorptiometry (DXA) on d26, d46, and d66 of postnatal life. Whole-body fat content increased (p=0.01; 11.2%), and whole-body lean tissue decreased (p=0.01; 2.6%) in IUPFTN versus IUTN and IUHS offspring. Whole-body composition was similar between IUHS and IUTN offspring. Epididymal fat pad weight increased (p=0.03; 21.6%) in IUPFTN versus IUHS offspring. In summary and in contrast to pigs, IUHS did not impact rodent body composition during this stage of growth; however, IUPFTN altered the future hierarchy of tissue accretion.

The objectives of this study were to examine the effects of porcine reproductive and respiratory syndrome virus (PRRSV) infection and vaccination on pig growth, dietary nutrient efficiency of utilization, manure output, and emissions of CO, CH, HS, NO, and NH gases from stored manure. Forty-eight pigs, aged 21 d at the start of the study, were subjected to 1 of 4 treatment combinations arranged in a 2 × 2 factorial design with main factors of PRRSV vaccination and PRRSV infection. Body weight, ADFI, manure output, and nutrient efficiency of utilization were assessed and gas emissions from stored manure were determined daily from 50 to 78 d of age and for 24 d after completion of the animal phase. Infection with PRRSV markedly reduced final BW, ADG, and ADFI ( < 0.01) and reduced efficiencies of ADF and ether extract utilization ( = 0.05 and = 0.02, respectively) regardless of vaccination status. No significant treatment effects were found on manure output, manure pH, efficiencies of lignin utilization, and N retention. Infecting pigs with PRRSV increased daily manure CO emission per pig ( = 0.01). There was an interaction between immunization and infection for NO per pig with manure from uninfected, vaccinated pigs producing as much as the manure from infected, vaccinated pigs whereas there was a difference by PRRSV infection state for nonvaccinated pigs. There were also interactions between treatments for HS and NO emissions per kilogram of manure volatile solids excreted ( = 0.01 and = 0.0001, respectively) with the same pattern as for NO per pig; that is, the vaccinated pigs had similar rates of emission regardless of infection state. Pigs infected with PRRSV increased NO nitrogen per kilogram of total N excreted compared with noninfected groups ( = 0.03). Collectively, these results indicated that PRRSV infection caused decreased growth rates and nutrient utilization efficiency and increased gas emissions from stored manure.

In utero heat stress (IUHS) negatively impacts postnatal development, but how it alters future body temperature parameters and energetic metabolism is not well understood. Future body temperature indices and bioenergetic markers were characterized in pigs from differing in utero thermal environments during postnatal thermoneutral (TN) and cyclical heat stress (HS) exposure. First-parity pregnant gilts ( = 13) were exposed to 1 of 4 ambient temperature (T) treatments (HS [cyclic 28°C to 34°C] or TN [cyclic 18°C to 22°C]) applied for the entire gestation (HSHS, TNTN), HS for the first half of gestation (HSTN), or HS for the second half of gestation (TNHS). Twenty-four offspring (23.1 ± 1.2 kg BW; = 6 HSHS, = 6 TNTN, = 6 HSTN, = 6 TNHS) were housed in TN (21.7°C ± 0.7°C) conditions and then exposed to 2 separate but similar HS periods (HS1 = 6 d; HS2 = 6 d; cycling 28°C to 36°C). Core body temperature (T) was assessed every 15 min with implanted temperature recorders. Regardless of in utero treatment, T increased during both HS periods ( = 0.01; 0.58°C). During TN, HS1, and HS2, all IUHS pigs combined had increased T ( = 0.01; 0.36°C, 0.20°C, and 0.16°C, respectively) compared to TNTN controls. Although unaffected by in utero environment, the total plasma thyroxine to triiodothyronine ratio was reduced ( = 0.01) during HS1 and HS2 (39% and 29%, respectively) compared with TN. In summary, pigs from IUHS maintained an increased T compared with TNTN controls regardless of external T, and this thermal differential may have practical implications to developmental biology and animal bioenergetics.

The objective of this study was to evaluate in cattle, the effects of acute exposure to a heat stress (HS) environment on the status of the pituitary (thyrotropin, TSH)-thyroid (thyroxine, T4)-peripheral tissue T4 deiodination (type 1 5'-deiodinase [D1]; triiodothyronine [T3]; reverse-triiodothyronine [rT3]) axis, and the further response of this pituitary-thyroid-peripheral tissue axis (PTTA) to perturbation caused by the induction of the proinflammatory innate immune state provoked by the administration of gram-negative bacteria endotoxin (lipopolysaccharide [LPS]). Ten steers (318 ± 49 kg body weight) housed in controlled environment chambers were subjected to either a thermoneutral (TN: constant 19°C) or HS temperature conditions (cyclical daily temperatures: 32.2°C-40.0°C) for a total period of 9 d. To minimize the effects of altered plane of nutrition due to HS, steers in TN were pair-fed to animals in HS conditions. Steers received 2 LPS challenges 3 d apart (LPS1 and LPS2; 0.2 μg/kg body weight, intravenously, Escherichia coli 055:B5) with the first challenge administered on day 4 relative to the start of the environmental conditioning. Jugular blood samples were collected at 0, 1, 2, 4, 7, and 24 h relative to the start of each LPS challenge. Plasma TSH, T4, T3, and rT3 were measured by radioimmunoassay. Liver D1 activity was measured in biopsy samples collected before the LPS1 (0 h) and 24 h after LPS2. Before the start of LPS1, HS decreased (P0.01 vs TN) plasma TSH (40%), T4 (45.4%), and T3 (25.9%), but did not affect rT3 concentrations. In TN steers, the LPS1 challenge decreased (P0.01 vs 0 h) plasma concentrations of TSH between 1 and 7 h and T4 and T3 at 7 and 24 h. In HS steers, plasma TSH concentrations were decreased at 2 h only (P0.05), whereas plasma T3 was decreased at 7 and 24 h (P0.01). Whereas plasma T4 concentrations were already depressed in HS steers at 0 h, LPS1 did not further affect the levels. Plasma rT3 concentrations were increased in all steers at 4, 7, and 24 h after LPS1 (P0.01). The patterns of concentration change of T4, T3, and rT3 during LPS2 mirrored those observed in LPS1; the responses in plasma TSH were of smaller magnitude than those incurred after LPS1. The LPS challenges reduced (P0.01) hepatic activity of D1 in all animals but no differences were observed between steers subjected to TN or HS environment. The data are consistent with the concept that acute exposure of cattle to a HS environment results in the depression of the pituitary and thyroid components of the PTTA, whereas a normal capacity to generate T3 from T4 in the liver is preserved. The data also suggest that LPS challenge further suppresses all components of the PTTA including liver T3 generation, and these PTTA perturbations are more pronounced in steers that encounter a HS exposure.

Heat stress (HS) negatively impacts several swine production variables, including carcass fat quality and quantity. Pigs reared in HS have more adipose tissue than energetically predicted, explainable, in part, by HS-induced hyperinsulinemia. Study objectives were to evaluate insulin’s role in altering fat characteristics during HS via feeding insulin-sensitizing compounds. Forty crossbred barrows (113 ± 9 kg BW) were randomly assigned to one of five environment by diet treatments: 1) thermoneutral (TN) fed ad libitum (TNAL), 2) TN and pair-fed (TNPF), 3) HS fed ad libitum (HSAL), 4) HS fed ad libitum with sterculic oil (SO) supplementation (HSSO; 13 g/d), and 5) HS fed ad libitum with dietary chromium (Cr) supplementation (HSCr; 0.5 mg/d; Kemin Industries, Des Moines, IA). The study consisted of three experimental periods (P). During P0 (2 d), all pigs were exposed to TN conditions (23 ± 3 °C, 68 ± 10% RH) and fed ad libitum. During P1 (7 d), all pigs received their respective dietary supplements, were maintained in TN conditions, and fed ad libitum. During P2 (21 d), HSAL, HSSO, and HSCr pigs were fed ad libitum and exposed to cyclical HS conditions (28 to 33 °C, 58 ± 10% RH). The TNAL and TNPF pigs remained in TN conditions and were fed ad libitum or pair-fed to their HSAL counterparts. Rectal temperature (T(R)), respiration rate (RR), and skin temperature (T(S)) were obtained daily at 0600 and 1800 h. At 1800 h, HS exposed pigs had increased T(R), RR, and T(S) relative to TNAL controls (1.13 °C, 48 bpm, and 3.51 °C, respectively; P < 0.01). During wk 2 and 3 of P2, HSSO pigs had increased 1800 h T(R) relative to HSAL and HSCr (~0.40 and ~0.42 °C, respectively; P ≤ 0.05). Heat stress decreased ADFI and ADG compared to TNAL pigs (2.24 vs. 3.28 and 0.63 vs. 1.09 kg/d, respectively; P < 0.01) and neither variable was affected by SO or Cr supplementation. Heat stress increased or tended to increase moisture content of abdominal (7.7 vs. 5.9%; P = 0.07) and inner s.c. (11.4 vs. 9.8%; P < 0.05) adipose depots compared to TNAL controls. Interestingly, TNPF pigs also had increased adipose tissue moisture content and this was most pronounced in the outer s.c. depot (15.0 vs. 12.2%; P < 0.01) compared to TNAL pigs. Heat stress had little or no effect on fatty acid composition of abdominal, inner, and outer s.c. adipose tissue depots. In summary, the negative effects of HS on fat quality do not appear to be fatty acid composition related, but may be explained by increased adipose tissue moisture content.

The detrimental effects of heat stress (HS) on animal productivity have been well documented. However, whether in utero HS interacts with a future thermal insult to alter tissue deposition during the finishing phase of pig growth is unknown. Study objectives were to compare the subsequent rate and quantity of whole-body tissue accretion in pigs exposed to differing in utero and postnatal thermal environments. Pregnant sows were exposed to thermoneutral (TN; cyclical 15°C nighttime and 22°C daytime; n = 9) or HS (cyclical 27°C nighttime and 37°C daytime; n = 11) conditions during their entire gestation. Twenty-four offspring from in utero TN (IUTN; n = 6 gilts and 6 barrows; 62.4 ± 0.7 kg BW) and in utero HS (IUHS; n = 6 gilts and 6 barrows; 61.9 ± 0.8 kg BW) were euthanized as part of an initial slaughter group (ISG). After the ISG, 48 pigs from IUTN (n = 12 gilts and 12 barrows; 66.1 ± 1.0 kg BW) and IUHS (n = 12 gilts and 12 barrows; 63.4 ± 0.7 kg BW) were exposed to constant HS (34.4 ± 1.8°C) or TN (22.7 ± 2.5°C) conditions until they reached 80.5 ± 1.5 kg BW, at which point they were sacrificed and their whole-body composition was determined. Homogenized carcasses were analyzed for N, crude fat, ash, water, and GE content. Data were analyzed using PROC MIXED in SAS 9.3. Rectal temperature and respiration rate increased during postnatal HS compared to TN (39.6 vs. 39.3°C and 92 vs. 58 breaths per minute, respectively; P < 0.01). Postnatal HS decreased (P < 0.01) feed intake (2.13 vs. 2.65 kg/d) and ADG (0.70 vs. 0.94 kg/d) compared to TN conditions, but neither variable was influenced by in utero environment. Whole-body protein and lipid accretion rates were reduced in HS pigs compared to TN controls (126 vs. 164 g/d and 218 vs. 294 g/d, respectively; P < 0.04). Independent of postnatal environments, IUHS reduced future protein accretion rates (16%; P < 0.01) and tended to increase lipid accretion rates (292 vs. 220 g/d; P < 0.07) compared to IUTN controls. The ratio of lipid to protein accretion rates increased (95%; P < 0.01) in IUHS pigs compared to IUTN controls. In summary, the future hierarchy of tissue accretion is altered by IUHS, and this modified nutrient partitioning favors adipose deposition at the expense of skeletal muscle during this specific phase of growth.

Heat stress (HS) and immune challenges negatively impact nutrient allocation and metabolism in swine, especially due to elevated heat load. In order to assess the effects of HS during Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) infection on metabolism, 9-wk old crossbred barrows were individually housed, fed ad libitum, divided into four treatments: thermo-neutral (TN), thermo-neutral PRRSV infected (TP), HS, and HS PRRSV infected (HP), and subjected to two experimental phases. Phase 1 occurred in TN conditions (22 °C) where half the animals were infected with PRRS virus (n = 12), while the other half (n = 11) remained uninfected. Phase 2 began, after 10 d with half of the uninfected (n = 6) and infected groups (n = 6) transported to heated rooms (35 °C) for 3 d of continuous heat, while the rest remained in TN conditions. Blood samples were collected prior to each phase and at trial completion before sacrifice. PPRS viral load indicated only infected animals were infected. Individual rectal temperature (T(r)), respiration rates (RR), and feed intakes (FI) were determined daily. Pigs exposed to either challenge had an increased T(r), (P < 0.0001) whereas RR increased (P < 0.0001) with HS, compared to TN. ADG and BW decreased with challenges compared to TN, with the greatest loss to HP pigs. Markers of muscle degradation such as creatine kinase, creatinine, and urea nitrogen were elevated during challenges. Blood glucose levels tended to decrease in HS pigs. HS tended to decrease white blood cell (WBC) and lymphocytes and increase monocytes and eosinophils during HS. However, neutrophils were significantly increased (P < 0.01) during HP. Metabolic flexibility tended to decrease in PRRS infected pigs as well as HS pigs. Fatty acid oxidation measured by CO(2) production decreased in HP pigs. Taken together, these data demonstrate the additive effects of the HP challenge compared to either PRRSV or HS alone.

Sex steroid hormones are used in the meat industry due to their ability to regulate muscle hypertrophy. However, the mechanisms underlying their action are not fully elucidated. Recent reports demonstrate that steroid hormones increase oxytocin (OXT) expression in skeletal muscle, indicating that OXT may play a role in satellite cell activity. This hypothesis was tested using steroid hormones (17β-estradiol [E2]; trenbolone acetate [TBA]), tamoxifen (TAM), OXT, and atosiban (A: OXT receptor inhibitor) applied to bovine satellite cells (BSCs) to investigate BSC regulation by OXT. Oxytocin alone increased fusion index (P 0.05) but not BSC proliferation. Oxytocin reduced (P0.05) apoptotic nuclei and stimulated migration rate (P0.05). Similarly, E2 and TBA increased (P0.05) BSC proliferation rate, fusion index, and migration and decreased (P0.05) apoptotic nuclei. 17β-Estradiol or TBA supplemented with A had lower (P0.05) BSC proliferation rate, fusion index, and migration and more (P0.05) apoptotic nuclei compared with E2 or TBA alone. Furthermore, OXT expression increased (P0.05) in E2 or TBA-treated proliferating BSC. Oxytocin, E2, and TBA increased (P0.05) MyoD and MyoG expression in proliferating BSC. During BSC differentiation, OXT expression increased (P0.05) with E2 or TBA treatments. MyoG expression increased (P0.05) in OXT, E2, and TBA compared with control. However, A, OXT + A, TAM, TAM + OXT, E2 + TAM, E2 + A, and TBA + A decreased (P0.05) MyoG expression during BSC differentiation. These results indicate that OXT is involved in steroid hormone-stimulated BSC activity.

Despite well-studied clinical manifestations, intracellular mechanisms of prolonged hyperthermic injury remain unclear, especially in skeletal muscle. Given muscle's large potential to impact systemic inflammation and metabolism, the response of muscle cells to heat-mediated injury warrants further investigation. We have previously reported increased activation of NF-κB signaling and increased NF-κB and AP-1-driven transcripts in oxidative skeletal muscle following 12 h of heat stress. The purpose of this investigation was to examine early heat stress-induced inflammatory signaling in skeletal muscle. We hypothesized that heat stress would increase NF-κB and AP-1 signaling in oxidative skeletal muscle. To address this hypothesis, 32 gilts were randomly assigned to one of four treatment groups (n = 8/group): control (0 h: 21°C) or exposed to heat stress conditions (37°C) for 2 h (n = 8), 4 h (n = 8), or 6 h (n = 8). Immediately following environmental exposure pigs were euthanized and the red portion of the semitendinosus muscle (STR) was harvested. We found evidence of NF-κB pathway activation as indicated by increased protein abundance of NF-κB activator IKK-α following 4 h and increased total NF-κB protein abundance following 6 h of heat stress. Heat stress also stimulated AP-1 signaling as AP-1 protein abundance was increased in nuclear fractions following 4 h of heat stress. Interleukin-6 protein abundance and activation of the JAK/STAT pathway were decreased in heat stressed muscle. These data indicate that heat stress activated inflammatory signaling in the porcine STR muscle via the AP-1 pathway and early activation of the NF-κB pathway. Published version

We have previously established that 24h of environmental hyperthermia causes oxidative stress and have implicated mitochondria as likely contributors to this process. Given this, we hypothesized that heat stress would lead to increased autophagy/mitophagy and a reduction in mitochondrial content. To address this hypothesis pigs were housed in thermoneutral (TN; 20 degrees C) or heat stress (35 degrees C) conditions for 1- (HS1) or 3- (HS3) days and the red and white portions of the semitendinosus collected. We did not detect differences in glycolytic muscle. Counter to our hypothesis, upstream activation of autophagy was largely similar between groups as were markers of autophagosome nucleation and elongation. LC3A/B-I increased 1.6-fold in HS1 and HS3 compared to TN (P < 0.05), LC3A/B-II was increased 4.1-fold in HS1 and 4.8-fold in HS3 relative to TN, (P < 0.05) and the LC3A/B-II/I ratio was increased 3-fold in HS1 and HS3 compared to TN suggesting an accumulation of autophagosomes. p62 was dramatically increased in HS1 and HS3 compared to TN. Heat stress decreased mitophagy markers PINK1 7.0-fold in HS1 (P < 0.05) and numerically by 2.4-fold in HS3 compared to TN and BNIP3L/NIX by 2.5-fold (P < 0.05) in HS1 and HS3. Markers of mitochondrial content were largely increased without activation of PGC-1 signaling. In total, these data suggest heat-stress-mediated suppression of activation of autophagy and autophagosomal degradation, which may enable the persistence of damaged mitochondria in muscle cells and promote a dysfunctional intracellular environment. USDA [2014-67015-21627, 20116700330007]; Wise Burrough's fund This project was supported by USDA grants 2014-67015-21627 (JTS) and 20116700330007 (LHB) and AB was supported by the Wise Burrough's fund.

Heat stress contributes to higher morbidity and mortality in humans and animals and is an agricultural economic challenge because it reduces livestock productivity. Redox balance and associated mitochondrial responses appear to play a central role in heat stress-induced skeletal muscle pathology. We have previously reported increased oxidative stress and mitochondrial content in oxidative muscle following 12h of heat stress. The purposes of this investigation were to characterize heat stress-induced oxidative stress and changes in mitochondrial content and biogenic signaling in oxidative skeletal muscle. Crossbred gilts were randomly assigned to either thermal neutral (21 degrees C; n=8, control group) or heat stress (37 degrees C) conditions for 2h (n=8), 4h (n=8), or 6h (n=8). At the end, their respective environmental exposure, the red portion of the semitendinosus muscle (STR) was harvested. Heat stress increased concentration of malondialdehyde (MDA) following 2 and 4h compared to thermal neutral and 6h, which was similar to thermal neutral, and decreased linearly with time. Protein carbonyl content was not influenced by environment. Catalase activity was increased following 4h of heat stress and superoxide dismutase activity was decreased following 6h of heat stress compared to thermal neutral conditions. Heat stress-mediated changes in antioxidant activity were independent of altered protein abundance or transcript expression. Mitochondrial content and mitochondrial biogenic signaling were similar between groups. These data demonstrate that heat stress caused a transient increase in oxidative stress that was countered by a compensatory change in catalase activity. These findings contribute to our growing understanding of the chronology of heat stress-induced intracellular dysfunctions in skeletal muscle. USDA grants [2014-67015-21627, 2011-6700330007] This work was supported by USDA grants 2014-67015-21627 (J. T. S.) and 2011-6700330007 (L. H. B.).